Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

Apurinic/apyrimidinic endonuclease 1(APE 1)is a multifunctional protein that plays important roles in DNA repair and regulation of gene expression.Because APE 1 is overexpressed in various cancers,it can serve as a cancer biomarker for aiding clinical diagnosis,guiding therapy,and monitoring prognosis.On this basis,a label-free fluorescent probe was designed based on the primer exchange reaction(PER)strategy for highly sensitive detection of APE 1 activity.In the absence of APE 1,the structure of catalytic hairpin(HP)was stable and could not form G-quadruplex.Therefore,the background fluorescence of this sensing system was very low due to the dissociation of thioflavin T(ThT).In the presence of APE 1,the apurinic/apyrimidinic(AP)site of HP was cleaved by APE 1 and a short nucleic acid fragment that acted as a primer to initiate PER was generated.After PER reaction,a large number of G-quadruplex were produced,which could specifically bind with ThT and resulted in significant increase of fluorescence signal.The combination of low background design of HP and PER amplification made this biosensor had high sensitivity with a detection limit(3σ)of 0.0008 U/mL.Furthermore,the primer sequence was directly generated by the cleavage of APE 1 without additional addition,which not only increased the specificity of the reaction,but also simplified the experiment procedure.Moreover,the use of label-free fluorescence signal reduced the cost of the experiment,and realized rapid detection of APE 1.Finally,this sensor was used to detect APE 1 in human serum samples with spiked recoveries of 91%-104%,proving great potential in study of biological enzyme.

OBJECTIVE: The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment. METHODS: Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ. RESULTS: Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein β1 (Hspb1) was the highest in the EA group compared to the model group. CONCLUSION EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.
Rats ; Male ; Animals ; Rats, Sprague-Dawley ; Electroacupuncture ; Proteomics ; Cognitive Dysfunction/therapy* ; Hippocampus

The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis. The single-cell RNA sequencing (scRNA-seq) analysis of the testis was performed to identify genes upregulated in spermatogonia. Using scRNA-seq analysis, we identified the spermatogonia upregulated gene origin recognition complex subunit 6 (Orc6), which is involved in DNA replication and cell cycle regulation; its protein expression in the human and mouse testis was detected by western blot and immunofluorescence. To explore the potential function of Orc6 in spermatogonia, the C18-4 cell line was transfected with control or Orc6 siRNA. Subsequently, 5-ethynyl-2-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, flow cytometry, and western blot were used to evaluate its effects on proliferation and apoptosis. It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells. Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated (Wnt)/ β-catenin signaling. Western blot revealed that the expression of β-catenin protein and its phosphorylation (Ser675) were significantly decreased when silencing the expression of ORC6. Our findings indicated that Orc6 was upregulated in spermatogonia, whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.

Objective:To estimate the incidence and mortality of corpus uteri cancer in China, 2015.Methods:Quality audit and evaluation of the data from 2015 cancer registration reported by 501 cancer registries were conducted, and 368 cancer registries were included in the analysis. The incidence rate and mortality rate of corpus uteri cancer were calculated according to the factors of the region (urban, rural, east, central, western), sex and age groups. The incidence and mortality of corpus uteri cancer with the 2015 population were estimated. Chinese standard population in 2000 and world Segi′s population were used for the calculation of age-standardized rates (ASR) of incidence and mortality.Results:In 2015, 368 cancer registries included in the analysis covered a total of 309 553 499 populations in China, accounting for 22.52% of the national population. It is estimated that there were about 68 900 new cases of corpus uteri cancer in 2015, the incidence rate was 10.28/10 5, age-standardized incidence rates by Chinese standard population (ASR China) and world standard population (ASR world) were 6.86/10 5 and 6.66/10 5, respectively. The incidence rate of urban area (11.35/10 5) was higher than that of rural area (8.90/10 5), and the incidence of eastern region (12.12/10 5) was higher than the central region (9.94/10 5) and the western region (8.25/10 5). It is estimated that in 2015, there were about 16 000 deaths of corpus uteri cancer, the mortality rate was 2.39/10 5, ASR China was 1.49/10 5, ASR world was 1.47/10 5. The mortality in urban areas (2.40/10 5) is close to rural areas (2.39/10 5); the mortality in central areas (2.55/10 5) was higher than the eastern areas (2.32/10 5) and the western areas (2.31/10 5). Conclusions:In China, the incidence of corpus uteri cancer is on the rise and has a trend of youth, the burden of disease is gradually increasing, which threatens the health of women. Targeted prevention and control measures should be carried out in the different regions.

Objective:To estimate the incidence and mortality of corpus uteri cancer in China, 2015.Methods:Quality audit and evaluation of the data from 2015 cancer registration reported by 501 cancer registries were conducted, and 368 cancer registries were included in the analysis. The incidence rate and mortality rate of corpus uteri cancer were calculated according to the factors of the region (urban, rural, east, central, western), sex and age groups. The incidence and mortality of corpus uteri cancer with the 2015 population were estimated. Chinese standard population in 2000 and world Segi′s population were used for the calculation of age-standardized rates (ASR) of incidence and mortality.Results:In 2015, 368 cancer registries included in the analysis covered a total of 309 553 499 populations in China, accounting for 22.52% of the national population. It is estimated that there were about 68 900 new cases of corpus uteri cancer in 2015, the incidence rate was 10.28/10 5, age-standardized incidence rates by Chinese standard population (ASR China) and world standard population (ASR world) were 6.86/10 5 and 6.66/10 5, respectively. The incidence rate of urban area (11.35/10 5) was higher than that of rural area (8.90/10 5), and the incidence of eastern region (12.12/10 5) was higher than the central region (9.94/10 5) and the western region (8.25/10 5). It is estimated that in 2015, there were about 16 000 deaths of corpus uteri cancer, the mortality rate was 2.39/10 5, ASR China was 1.49/10 5, ASR world was 1.47/10 5. The mortality in urban areas (2.40/10 5) is close to rural areas (2.39/10 5); the mortality in central areas (2.55/10 5) was higher than the eastern areas (2.32/10 5) and the western areas (2.31/10 5). Conclusions:In China, the incidence of corpus uteri cancer is on the rise and has a trend of youth, the burden of disease is gradually increasing, which threatens the health of women. Targeted prevention and control measures should be carried out in the different regions.

Objective: To determine the prevalence and determinants of folic acid (FA) supplementation in Chinese couples planning for pregnancy and in women during early pregnancy. Methods: This was a cross-sectional study based on the Shanghai PreConception Cohort (SPCC) study. Data on FA supplementation and socio-demographic features were collected using questionnaires. Couples visiting clinics for pre-pregnancy examination and pregnant women at < 14 gestational weeks were recruited in Shanghai, China, between March 2016 and September 2018. Results: Among the pregnancy planners, 42.4% (4,710/11,099) women and 17.1% (1,377/8,045) men used FA supplements, while 93.4% (14,585/15,615) of the pregnant women used FA supplements. FA supplement use was higher in female pregnancy planners who were older ( : 1.13, 95% : 1.08-1.18), had higher education ( : 1.71, 95% : 1.53-1.92), and were residing in urban districts ( : 1.06, 95% : 1.01-1.11) of FA supplementation; female pregnancy planners with alcohol consumption ( : 0.95, 95% : 0.90-0.99) had lower odds of FA supplementation. In early pregnancy, women with higher educational level ( : 1.04, 95% : 1.03-1.06), who underwent pre-pregnancy examination ( : 1.02, 95% : 1.01-1.03) had higher odds of using an FA supplement; older aged ( : 0.99, 95% : 0.98-0.99), and multigravida ( : 0.97, 95% : 0.96-0.98) had lower odds of FA supplementation. Conclusion Although the majority of pregnant women took FA supplements, more than half of the women planning for pregnancy did not. Urgent strategies are needed to improve pre-conception FA supplementation.
Adult ; China ; Cohort Studies ; Cross-Sectional Studies ; Diet ; Dietary Supplements ; analysis ; Female ; Folic Acid ; administration & dosage ; Humans ; Male ; Pregnancy ; Surveys and Questionnaires ; Vitamin B Complex ; administration & dosage ; Young Adult

Amyloid ; Amyloid Neuropathies, Familial/genetics* ; China ; Humans ; Mutation ; Prealbumin/genetics*

Objective　Dendritic cells (DCs), helper T cells 17 (Th17) and regulatory T cells (Treg) are closely related to the pathogenesis of chronic obstructive pulmonary disease (COPD). This study aimed to investigate the changes of Th17- and Treg-related cytokines in the bronchoalveolar lavage fluid (BALF) of COPD mice after DC-based adoptive immunotherapy with over-expressed suppressor of cytokine signaling protein 1 (SOCS1) and provide some new ideas for the treatment of COPD.　Methods　A total of 48 male C57BL/6 mice were randomly divided into 5 groups: healthy control, COPD model control, immature DC (imDC), DC-SOCS1 1×106, and DC-SOCS1 2×106. The healthy controls were exposed to air and fed normally, the COPD model controls injected with normal saline at 0.5 mL/ on the first day of modeling by fumigation, the mice of the imDC group injected via the tail vein with 1 ×106 imDCs, and those of the DC-SOCS1 groups injected with 1 ×106 or 2 ×106 DCs with over expressed SOCS1, all via the tail vein on the 1st and 7th day of modeling. Then the lung tissues were collected from the mice for preparation of paraffin sections and HE staining, and ELISA was employed for determination of the levels of Th17-related IL-17 and IL-23 and Treg-related IL-10 and TGF-β in the BALF of the model mice.　Results　Compared with the COPD model controls, the mice in the imDC, DC-SOCS1 1×106 and DC-SOCS1 2×106 groups showed significantly decreased levels of IL-17 on the 1st day ([78.87 ± 1.08] vs [46.46 ± 0.77], [34.09 ± 3.98] and [24.12 ± 0.57] pg/mL, P < 0.05) and 7th day after modeling ([78.87 ± 1.08] vs [55.69 ±0.35], [35.65 ± 0.54] and [27.00 ± 0.58] pg/mL, P < 0.05), and IL-23 on the 1st day ([200.62 ± 0.65] vs [150.19 ± 0.53], [121.09 ± 0. 53] and [70.21 ± 0.91] pg/mL, P < 0.05) and 7th day ([200.62 ± 0.65] vs [167.70 ± 1.73], [136.34 ± 0.90] and [99.35 ± 1.83] pg/mL, P < 0.05), but remarkably increased levels of IL-10 on the 1st day ([39.46 ± 3.88] vs [50.74 ± 1.77], [58.71 ± 3.84] and [70.12 ± 2.62] pg/mL, P < 0.05) and 7th day ([39.46 ± 3.88] vs [44.56 ± 2.63], [54.78 ± 1.43] and [63.00 ± 2.57] pg/mL, P < 0.05), TGF-β on the 1st day ([24.98 ± 0.43] vs [36.46 ± 0.98], [42.40 ± 0.62] and [50.55 ± 0.53] pg/mL, P < 0.05) and 7th day ([24.98 ± 0.43] vs [33.27 ± 0.92], [40.12 ± 0.83] and [44.98 ± 0.52] pg/mL, P < 0.05). The contents of IL-17 and IL-23 were markedly lower while those of IL-10 and TGF-β higher in the DC-SOCS1 1×106 than in the imDC group (P < 0.05), and the levels of the former two significantly higher and those of the latter two lower in the DC-SOCS1 2×106 than in the DC-SOCS1 1×106 group (P < 0.05).　Conclusion　Transfusion of DCs with over-expressed SOCS1 can inhibit the secretion of Th17-related cytokines in COPD, and the effect is better than that of imDCs alone and related to the concentration and time.